LOCUS       V028813                10003 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028813
VERSION     V028813
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        9..235
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             236..416
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    463..588
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1081..1314
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1499..1543
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1810..1926
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2130..3312
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3335..4786
                     /label="Slc16a7(NM_011391)"
                     /note="Slc16a7(NM_011391)"
                     /gene="Slc16a7"
     misc_feature    4835..5408
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5442..5840
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5855..6443
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6530..6763
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6835..6956
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6996..7131
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(7152..7170)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7180..7196)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      7338..7793
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7819..7923
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7924..8784
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      8955..9543
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9831..9852
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9867..9897
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9905..9921
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     9929..9945
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9966..9984
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
       61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
      121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
      181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
      541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
      601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
      661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
      721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
      781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
      841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
      901 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
      961 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1021 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1081 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1141 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1201 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1261 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1321 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1381 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     1441 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     1501 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     1561 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     1621 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     1681 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     1741 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     1801 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     1861 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa
     1921 aattttatcg atgcctcccc gtcaccaccc cccccaaccc gccccgaccg gagctgagag
     1981 taattcatac aaaaggactc gcccctgcct tggggaatcc cagggaccgt cgttaaactc
     2041 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc
     2101 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca
     2161 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     2221 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2281 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2341 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     2401 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt
     2461 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     2521 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     2581 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     2641 ttttgatgat atcctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     2701 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     2761 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     2821 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     2881 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     2941 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     3001 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     3061 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     3121 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     3181 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     3241 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     3301 caggtgtcgt gaaaactacc cctctagagc caccatgcca tcagagcctt ccgcgccgct
     3361 tccacaacca cttcctcccg acggagggtg gggctgggtc gtagtctgtg cgtcctttat
     3421 ctccattgga ttttcatatg ccttccccaa agctgtcaca gtattcttca aagacatcca
     3481 ggaaatattc aacaccacct ccagtcagat cgcttggata tcgtccatca tgctagctgt
     3541 catgtatgcg ggaggtccca tcagtagtgt gttggtgaat aactatggca gccggcctgt
     3601 ggtgatagtg ggaggactat tatgctgcat tggaatgatc ttggcttctt acagcaacag
     3661 cgtgatagag ctttatctca ccgttggctt cattggaggt ttaggattag cattcaacct
     3721 gcaaccagcc ttaacaataa ttggaaaata cttctataga agacgaccac ttgcaaatgg
     3781 ctgcgccatg gctggaagtc ctgttttctt aagcacactg gctcctttca atcagtacct
     3841 atttaacaac tatggctgga aggggagttt tttgattttg ggaggtatat ttttgcactc
     3901 ctgtgtggct gggtgcctca tgagacctgt tgggccaagt ccaaatacta agaagtctaa
     3961 aagtaaggtt ggctcaagac atgattcaac tctgaagaaa gccagtaagg tatcaacagc
     4021 acaaaaggtt aataggtttt tagatttcag cctttttatg catagaggat ttttgatcta
     4081 cctatctgga aacgtcatat tgtttttagg gatttttgcc cctattatat tcttggctca
     4141 gtacgctaaa cacataggag tggatgacta taactcagct tttttactgt ctgtgatggc
     4201 ttttattgat atgtttgctc ggccttctgt aggattaata gccaacacta gcttaatccg
     4261 tccacgaatc cagtacctct tcagctctgc aatcatcttt actggcatat gccatctctt
     4321 atgcccccta gcaactacct actcagcttt ggtggtctac gttgtatttt ttggattggg
     4381 atttggaagt attagcagtc tcctctttga atgccttatg gacattgttg gagctaccag
     4441 gttctccagt gctgtgggac tcacaacaat cgtggagtgt tgtccagttc ttttcggccc
     4501 tcctctggct ggtaaattgc ttgatataac tggagaatat aaatacctgt acatagccag
     4561 cggaacagtc gtgttagtat caggtactta cctcctgatt ggcaatgcta taaactatag
     4621 acttctcgac aaggaaagga agagagagaa ggcaaagaag aagaagtcag cttcacacgc
     4681 gtccagagaa atggaggccc tgaacagatc taaacaggac gaagttactg tcaaagcttc
     4741 aaacgcacac aatcctccct cagacaggga caaagagagt aatatttaag gatccacgcg
     4801 tctccggcct agggataaca gggtaatccg ctagcccctc tccctccccc ccccctaacg
     4861 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     4921 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     4981 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     5041 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     5101 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     5161 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     5221 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     5281 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     5341 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     5401 gatgataata tggccacact agagatccac cggtcgccac catggccaag cctttgtctc
     5461 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     5521 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     5581 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     5641 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     5701 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     5761 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     5821 gttatgtgtg ggagggctaa gcggccgcgt cgacaatcaa cctctggatt acaaaatttg
     5881 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5941 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6001 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6061 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6121 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6181 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6241 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     6301 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6361 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6421 ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacctttaag accaatgact
     6481 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6541 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     6601 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6661 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6721 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     6781 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6841 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6901 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6961 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     7021 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     7081 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     7141 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     7201 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     7261 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     7321 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     7441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     7501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     7801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7861 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7921 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7981 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     8041 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     8101 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     8161 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     8221 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     8281 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     8341 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     8401 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     8461 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     8521 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8581 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8641 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8701 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8761 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8821 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8881 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8941 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     9001 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     9061 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     9121 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     9181 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     9241 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     9301 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     9361 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     9421 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     9481 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     9541 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     9601 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     9661 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     9721 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     9781 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     9841 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     9901 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     9961 cgcgcaatta accctcacta aagggaacaa aagctggagc tgc
//
